Immunologic membrane markers have been utilized for the in situ identification of lymphoreticular cells within lymphoreticular tissues and in inflammatory and neoplastic lymphoreticular infiltrates of the skin. Frozen sections were layered with untreated sheep red blood cells (SRBC) for the detection of the E receptor on T cells, SRBC coated with 7S antibody for the detection of the Fc receptor (IgGEA) and SRBC coated with 19S antibody and complement (IgMEAC) for the detection of the C3 receptor. In normal lymphoreticular tissue, IgGEA selectively bound to areas colonized by macrophages, IgMEAC to B-dependent areas, whereas E showed no adherence. In chronic lymphatic leukemia, a predominant and selective adherence of IgMEAC was found in lymphoreticular organs which supports the B-cell nature of the neoplastic cells; in leukemic reticuloendotheliosis a selective IgMEAC adherence was observed and this favors a monocytoid origin of the hairy cells. Adherence of any indicator cell was practically absent from skin infiltrates of mycosis fungoides, which represents an additional piece of evidence for the T-cell nature of this disease process. In psoriasis, a considerable binding of IgGEA was observed, whereas IgMEAC did not adhere at all. This result indicates that the lymphocytic part of the infiltrate in this disease consists mainly of T cells.

Humans↗

PubMed1977

A simplified one-step procedure for the simultaneous determination of complement receptor lymphocytes and lymphocytes with membrane-bound immunoglobulins.

A simple one-step procedure for the demonstration of complement receptor lymphocytes (CRL) with complement-coated bacteria (BC) as indicator particles is described. With this assay the percentage of CRL in normal peripheral blood ranged between 6 and 21% (mean 11%). In a separately performed combined assay for lymphocytes with membrane-bound immunoglobulins (M-Ig) and lymphocytes, which form rosettes with complement-coated bacteria (BC-RFC), four different fractions of lymphocytes could be detected: (M-Ig+-BC+, M-Ig+-BC-,M-Ig--BC+, M-Ig--BC-). These results suggest that the subpopulation of lymphocytes with complement receptor sites overlaps with, but is not totally identical with the lymphocyte subpopulation bearing Ig on the surface.

Adult↗

PubMed1977

Receptors for IgM-coated erythrocytes on chronic lymphatic leukemia cells.

Our experiments show that lymphocytes of CLL patients, having typical B cell characteristics, form rosettes with IgM-coated bovine erythrocytes. Of 18 investigated patients, 3 to 78% (mean 29%) of the isolated lymphocytes reacted with EA-IgM. With mixed rosette assays. EA-IgM bound to cells bearing receptors for IgG as well, but not receptor-bearing lymphocytes. Rosette formation could be completely blocked by addition of IgM at concentrations as low as 0.17 mg/ml. Ten milligrams per milliliter of aggregated human IgG had no effect on the rosette formation with EA-IgM but completely abolished the binding of EA-IgG. Adult human or rabbit serum blocked the EA-IgM binding, whereas cord blood serum and FCS had no effect. These inhibition data indicate that EA-IgM binding does not occur via a somewhat altered IgG-Fc receptor but reacts with different membrane structures. That EA-IgM receptor can be cleaved off with trypsin and can be reconstituted after overnight cultivation, also supports this viewpoint. In contrast to the situation in normal subjects, in CLL patients the IgM receptors are demonstrable before overnight cultivation and are found on cells with B cell characteristics.

Animals↗

PubMed1977

The development of lymphocytes with T- or B-membrane determinants in the human foetus.

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Antibody Specificity↗

PubMed1976

[T-Lymphocyte shifts in patients with melanoma and bronchogenic carcinoma (author's transl)].

Lymphocyte subpopulations were investigated in 32 patients with malignant melanoma, 25 patients with bronchogenic carcinoma and 59 control subjects. Whereas the determination of membrane immunoglobulin-bearing lymphocytes and of complement receptor-bearing lymphocytes gave comparable results in the tumour patient and control group, significant differences were found in the T-lymphocyte population using the sheep red blood cell (SRBC) assay. The so-called "active" Wybran rosette test, which characterizes a T-cell subpopulation with an especially avid receptor for SRBC, gave significantly lower results in patients with melanoma (25% control 35%, p less than 0.01) and bronchogenic carcinoma (21% control 35% p less than 0.01). Determination of the total T-cell population using the Jondal rosette assay gave significantly lower values in the patients with melanoma (52%, control 63%, p less than 0.01), but not in those with lung cancer (65%). Low rosette values were detected even at low histological invasion levels (classified according to Clark) in the patients with melanoma. No correlation was found between the invasion level and the percentage of rosette-forming cells. The significance of these findings and the value of the rosette assay in the assessment of the immunological reactivity of tumour patients is discussed.

Adenocarcinoma↗

PubMed1976

[Developments in immunofluorescence (author's transl)].

This report presents a review of recent developments in immunofluorescence and their resultant applications. After a short introduction to the basic principles of immunofluorescence, this study deals with newer developments in utilizable fluorochromes, new methods in the production of antisera and fluorochrome-labelled conjugates and recent improvements in the optical system. In particular, the necessity of suitable characterization and examination in regard to the quality of the conjugates is described in detail. A separate chapter has been devoted to the rapidly-developing quantitative immunofluorescence technique and its application.

Antibodies↗

PubMed1976

An immunofluorescence procedure for the demonstration of antibodies to native, double-stranded DNA and of circulating DNA-anti-DNA complexes.

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Antibodies↗

PubMed1976

[Microfluorometric demonstration of thyroglobulin antibodies with the defined antigen substrate spheres (DASS) system].

Previous investigations showed that quantitative immunofluorescence using antigens covalently bound to agarose particles represents a reproducible and sensitive assay for antibodies in experimental antisera. In this paper data are presented which show that also thyroglobulin antibodies in patients sera can be demonstrated using the defined antigen substrate spheres (DASS) system with thyroglobulin as antigen. Sera of 45 patients with various thyroid diseases were investigated for the presence of thyroglobulin antibodies using passive haemagglutination and the quantitative immunofluorescence technique. Comparable results were obtained with both techniques. Advantages and disadvantages of both methods are discussed.

Autoantibodies↗

PubMed1976

B- and T-cell determination in ALL.

The influence of different cytostatic treatment schemes on T- and B-cell behaviour has been studied in children with ALL and UAL. Under these treatment schemes the percentage of B-cells decreased, while the percentage of T-cells remained within normal limits. In long-term remissions, we found normal values. In the first attack of ALL before therapy, a different behaviour of lymphoid cells was found. One group of patients had no lymphoid cells with membrane-bound Ig and no rosette forming cells; a second group had mainly T-cells, and a third group showed normal values of lymphoid cells with membrane-bound Ig, but the ability of rosette formation was slightly diminished. Whether this membrane-bound Ig is really cell-produced or only attached to the cell-membrane (blocking antibodies?) is discussed. Capping and pitching behaviour was observed. There were no differences in cell-membrane fluidity as compared with the normal controls.

Adolescent↗